Which of these methods is appropriate for making a mouse model for cystic fibrosis, which is a recessive disease caused by various loss-of-function mutations in the CFTR gene? Select all that apply. CFTR gene inactivated by a marker gene is introduced to embryonic stem cells CRISPR that knocks in a mutant copy of the gene to a mouse genome Pronuclear injection of a mutant copy of the gene to a mouse zygote RNAI that knocks down the wild-type gene expression in a mouse genome CRISPR that knocks out the wild-type gene in a mouse genome O O O 0 0
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- Mouse models for human genetic diseases are potentially powerful tools to help geneticists understand thecause of the aberrant phenotypes and develop newtherapeutic measures. However, such mice are not always as useful to investigators as it might seem at firstglance. Suppose that you have a mouse knockoutmodel for a human disease caused by homozygosityfor a null allele of a gene. Discuss how the followingsituations might complicate investigations of the human disease based on this mouse model.a. Mice have a shorter life span than humans.b. Mice homozygous for certain knockout mutationsdie in utero.c. Mouse genomes may have additional copies of thegene whose mutation causes the disease in humans.d. Mice from different inbred lines homozygous forthe same gene knockout vary in the penetrance andexpressivity of the phenotype.e. Manipulations to create the knockout mouse, suchas the presence of a drug resistance gene that allowsthe selection of cells containing the knockout (seeFig. 18.9),…Mouse models for human genetic diseases are potentially powerful tools to help geneticists understand thecause of the aberrant phenotypes and develop newtherapeutic measures. However, such mice are not always as useful to investigators as it might seem at firstglance. Suppose that you have a mouse knockoutmodel for a human disease caused by homozygosityfor a null allele of a gene. Discuss how the followingsituations might complicate investigations of the human disease based on this mouse model.a. Mice have a shorter life span than humans.b. Mice homozygous for certain knockout mutationsdie in utero.c. Mouse genomes may have additional copies of thegene whose mutation causes the disease in humans.If you wanted to make a mouse model for any of the following human genetic conditions (a–d), indicate which of thefollowing types of mice (i–vi) would be useful to your studies. If more than one answer applies, state which type ofmouse would most successfully mimic the human disease:(i) transgenic mouse overexpressing a normal mouse protein; (ii) transgenic mouse expressing normal amounts of amutant human protein; (iii) transgenic mouse expressing adominant negative form of a protein; (iv) a knockout mouse;(v) a conditional knockout mouse; and (vi) a knockin mousein which the normal allele is replaced with a mutant allelethat is at least partially functional. In all cases, the transgeneor the gene that is knocked out or knocked in is a form of thegene responsible for the disease in question.a. Marfan syndrome (a dominant disease caused byhaploinsufficiency for the FBN1 gene);b. A dominantly inherited autoinflammatory diseasecaused by a hypermorphic missense mutation in thegene PLCG2;c.…
- discuss the engineered embryronic stem cell method of obtaining gain of function transgenic miceIn the module, you have learned about P-element mediated transgenesis in Drosophila and the concept of using transgenes to rescue mutant phenotypes. In the figure below, you will see a wild type fly with its natural eye colour and three mutants with their eye colours changed to vermillion, white and rosy, respectively. A schematic of P-element mediated transgenesis (as shown in the lectures) is also included in the figure. Please inspect the schematic carefully and choose which of the following statements is true: I. Injection of the white experimental transgene into the vermillion mutant embryo will not change the vermillion mutant phenotype II. Injection of the white experimental transgene in the rosy mutant embryo will change rosy eye colour to red (wild type) III. Injection of the white experimental transgene in the white mutant embryo will not change the white mutant phenotype IV. Injection of the white experimental transgene in the rosy mutant…The researcher above decides to test whether any gene functional classes or pathways are enriched among the set of genes she has found to be differentially expressed between the treated and control mice. Provide the name of one online tool that can be used for this purpose and explain what is meant by the background gene set in a gene set enrichment analysis.
- You are interested in finding out in which organ GTF2H5 is highly expressed experimentally. Describe a method in recombinant DNA technology to measure gene expression, explain your rationale, and step by step, what are needed for this method to workWoolly mammoths have been extinct for about 4,000 years, but we often find their well-preserved remains in Siberian permafrost. Research groups are now planning to use SCNT (Somatic Cell Nuclear Transfer) to resurrect these huge elephant-like mammals. No mammoth eggs have been recovered yet, so elephant eggs would be used instead. An elephant would also be the surrogate mother for the resulting embryo. The researchers may try a modified SCNT technique used to clone a mouse that had been dead and frozen for sixteen years. Ice crystals that form during freezing break up cell membranes, so cells from the frozen mouse were in bad shape. Their DNA was transferred into donor mouse eggs, and cells from the resulting embryos were fused with undifferentiated mouse cells. Four healthy clones were born from the hybrid embryos. What are some of the pros and cons of cloning an extinct animal?Scientists carried out a microarray analysis to compare the gene expression of normal pancreatic cells to that of cancer cells from a person with pancreatic cancer. The scientists labeled the cDNA from the normal pancreatic cells with green fluorescent nucleotides. They labeled the cDNA from the cancer cells with red fluorescent nucleotides. The two cDNAs were mixed and allowed to hybridize to a microarray. Less p53 activity is found in cancer pancreatic cells than normal cells. What color would the spot for the p53 gene be on the microarray? Red Green Yellow Black
- You have a mouse model that is a homozygous knockout mutant for the prnp gene (Prnp protein is not produced in these mice). Describe what a prion is, describe how prions cause disease and, describe what would happen if you infected your prnp knockout mouse model with a prion. PLEASE ANSWER ALL PARTS TO THIS QUESTION AND PLEASE TYPE YOUR ANSWERThe TBX20 transcription factor is important for the developmentof heart tissue. Deletion of the Tbx20 gene in mice results in poorheart development and the death of mice well before birth. Tobetter understand how TBX20 regulates heart development ata genetic level, Sakabe et al. (2012. Hum. Mol. Genet. 21:2194–2204) performed a transcriptome analysis in which they comparedthe levels of all mRNAs between heart cells from wild-typemice and mice with Tbx20 deleted. This study concluded that TBX20 acts as an activator ofsome genes but a repressor of other genes in cardiac tissue.How might a single transcription factor have oppositeeffects on the transcription of different genes?Two genes associated with breast cancer, BRCA1 and BRCA2, were discovered in 1994 and 1995, respectively, and shortly thereafter, were patented by Myriad Genetics, a company based in Utah. Under the patents, testing for mutations in these genes could only be performed by Myriad, at costs from 300 to 3,000. Myriad also patented the process of analyzing the results of such tests, preventing anyone who obtains the sequence of their BRCA genes by other means (which itself would probably be patent infringement) from interpreting the information. The idea that genes can be patented has been a contentious issue from the beginning. Patents are not granted for products of nature, meaning that genes inside the body are not patentable, but biotech companies successfully argued that by removing a gene from the human body, purifying it, and then obtaining its DNA sequence, they created something not found in nature, and which is therefore a patentable invention. The U.S. Patent Office found the argument persuasive, but opponents argue that genes are parts of our bodies and can be identified but not invented. Biotech companies argue that without the protection offered by patents, they would have no incentive for research and development of diagnostic tests. In Europe, patents for BRCA1 and BRCA2 were revoked in 2004 because they did not meet the standards for a patent. After more than a decade of legal disputes, the patents were partially restored in 2008 on a very restricted basis. In the United States, a lawsuit, focused on the patents for the BRCA genes, was filed in May 2009. The suit challenges the basic idea that genes are patentable. In November 2009, the judge ruled that the lawsuit can proceed, and the case is moving forward. In March 2010, a federal court invalidated Myriad Genetics patent on these genes. In August 2011, the U.S. Court of Appeals reversed the lower courts decision and ruled that gene sequences isolated from cells are not a product of nature and are therefore patentable. The case went to the U.S. Supreme Court, which ordered the appeals court to reconsider the case. The Federal Appeals Court did not change its decision, and the case once again, went to the U.S. Supreme Court. A unanimous decision in June 2013 invalidated Myriads patents on the basis that isolating a gene from nature does not make it patentable. This is a landmark decision on gene patenting with widespread ramifications for the biotechnoloogy industry. Will this decision reduce the incentives for companies to invest in new diagnostic tests that would be used by cancer victims or those with serious genetic disorders?